Methods and compositions for impairing multiplication of HIV-1

Abstract

A composition which elicits antibodies to multiple known variants of Tat protein of HIV-1 of both the B and non-B clades contains the peptide R1-Asp-Pro-Asn-Leu-Asp-Pro-Trp-Asn-R2 SEQ ID NO: 23, and preferably an additional at least two variants of a peptide or polypeptide of the formula: R1-Asp-Pro-Y7-Leu-Glu-Pro-Trp-Z12-R2 SEQ ID NO: 8. In this composition, at least one of the two variants contains Arg at Y7 and Lys at Z12, and in at least a second of the two variants Y7 is Asn and Z12 is Asn. Vaccinal and pharmaceutical compositions can contain one or more such peptides associated with carrier proteins, associated in multiple antigenic peptides, or as part of recombinant proteins. Diagnostic compositions and uses are described for assessing the immune status of vaccinated patients.

What is claimed is:  
     
       1. A composition comprising a peptide or polypeptide of the formula R1-Asp-Pro-Asn-Leu-Asp-Pro-Trp-Asn-R2 SEQ ID NO: 8, 
       wherein R1 is selected from the group consisting of hydrogen, a lower alkyl, a lower alkanoyl, and a sequence of between 1 to about 5 amino acids, optionally substituted with a lower alkyl or lower alkanoyl; and  
       wherein R2 is selected from the group consisting of a free hydroxyl, an amide, and a sequence of one or up to about 5 additional amino acids, optionally substituted with an amide.  
     
     
       2. The composition according to  claim 1  wherein R1 is Val, resulting in the sequence Val-Asp-Pro-Asn-Leu-Asp-Pro-Trp-Asn of SEQ ID NO: 8. 
     
     
       3. The composition according to  claim 1  wherein R1 is X 2 -Pro-Val, wherein X 2  is selected from the group consisting of Glu and Asp, optionally substituted with a lower alkyl or lower alkanoyl, resulting in the sequence X 2 -Pro-Val-Asp-Pro-Asn-Leu-Asp-Pro-Trp-Asn SEQ ID NO: 38. 
     
     
       4. The composition according to  claim 1  wherein R2 is -His-Pro-Gly-Ser-, resulting in the sequence Val-Asp-Pro-Asn-Leu-Asp-Pro-Trp-Asn-His-Pro-Gly-Ser SEQ ID NO:16, wherein said carboxy-terminal Ser is optionally substituted with an amide. 
     
     
       5. The composition according to  claim 1  wherein said peptides or polypeptides are produced synthetically. 
     
     
       6. The composition according to  claim 1  wherein said peptides or polypeptides are produced recombinantly. 
     
     
       7. The composition according to  claim 1  wherein one or more of said peptides is expressed as a synthetic peptide coupled to a carrier protein. 
     
     
       8. The composition according to  claim 1  wherein one or more of said peptides is expressed as a multiple antigenic peptide, optionally coupled to a carrier protein. 
     
     
       9. The composition according to  claim 1  wherein one or more of the selected peptides is expressed within a recombinantly produced protein, optionally fused in frame with a carrier protein. 
     
     
       10. The composition according to any of claims  7 - 9 , wherein said carrier protein is selected from the group consisting of an  E. coli  DnaK protein, a GST protein, a mycobacterial heat shock protein 70, a diphtheria toxoid, a tetanus toxoid, a galactokinase, an ubiquitin, an α-mating factor, a β-galactosidase, and an influenza NS-1 protein. 
     
     
       11. A pharmaceutical composition comprising a composition of  claim 1 , a pharmaceutically acceptable carrier and an optional adjuvant. 
     
     
       12. The composition according to  claim 1 , further comprising at least two variants of a peptide or polypeptide of the formula selected from the group consisting of: 
       R1-Asp-Pro-Y 7 -Leu-Glu-Pro-Trp-Z 12 -R2 SEQ ID NO: 8,  
       wherein Y 7  is selected from the group consisting of Arg, Lys, Ser and Asn;  
       wherein Z 12  is selected from the group consisting of Lys and Asn;  
       wherein R1 is selected from the group consisting of hydrogen, a lower alky, a lower alkanoyl, and a sequence of between 1 to about 5 amino acids, optionally substituted with a lower alkyl or lower alkanoyl; and  
       wherein R2 is selected from the group consisting of a free hydroxyl, an amide, and a sequence of one or up to about 5 additional amino acids, optionally substituted with an amide, and  
       wherein at least one said variant is  
       R1-Asp-Pro-Arg-Leu-Glu-Pro-Trp-Lys-R2 SEQ ID NO: 17, and the other said variant is  
       R1-Asp-Pro-Asn-Leu-Glu-Pro-Trp-Asn-R2 SEQ ID NO: 18.  
     
     
       13. The composition according to  claim 12 , further comprising a peptide in which R1 is Val. 
     
     
       14. The composition according to  claim 12 , further comprising a peptide in which R1 is X 2 -Pro-Val, wherein X 2  is selected from the group consisting of Glu and Asp, optionally substituted with a lower alkyl or lower alkanoyl. 
     
     
       15. The composition according to  claim 12  further comprising a peptide wherein R2 is -His-Pro-Gly-Ser-, wherein said carboxy-terminal Ser is optionally substituted with an amide. 
     
     
       16. The composition according to  claim 12  comprising one or more additional sequences selected from the group consisting of: 
       R1-Asp-Pro-Lys-Leu-Glu-Pro-Trp-Lys-R2 SEQ ID NO: 19  
       R1-Asp-Pro-Lys-Leu-Glu-Pro-Trp-Asn-R2 SEQ ID NO: 22  
       R1-Asp-Pro-Ser-Leu-Glu-Pro-Trp-Lys-R2 SEQ ID NO: 20  
       R1-Asp-Pro-Ser-Leu-Glu-Pro-Trp-Asn-R2 SEQ ID NO: 24 and  
       R1-Asp-Pro-Asn-Leu-Glu-Pro-Trp-Lys-R2 SEQ ID NO: 21,  
       wherein R1 is selected from the group consisting of hydrogen, a lower alkyl, a lower alkanoyl, and a sequence of between 1 to about 5 amino acids, optionally substituted with a lower alkyl or lower alkanoyl; and  
       wherein R2 is selected from the group consisting of a free hydroxyl, an amide, and a sequence of one or up to about 5 additional amino acids, optionally substituted with an amide.  
     
     
       17. The composition according to  claim 16 , further comprising at least seven of said amino acid sequences.